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Consideration of the chemistry and biology of H2O2in cell culture:
• H2O2 has a relatively long half-life in solution and can diffuse in/out of the cell.
• Some compounds are easily oxidized in solution by air to produce H2O2 through a cell-independent mechanism.
• Several enzymes in the cell (e.g., monoamine oxidases (MAOs), D-amino acid oxidase, demethylases, etc.)directly produce H2O2 as a byproduct of their enzymatic reaction.
• Compounds, such as menadione and rotenone, that interfere with the electron transport chain of the mitochondria can induce H2O2 generation through the action of superoxide dismutase.
• Considerations important in determining a detection method for H2O2
• Ease of use
• Minimal false positive hits in the presence of library test compounds
• Adequate detection sensitivity
• Appropriate pH, temperature and buffer

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